Innovation
Focusing on the research and development and application of isothermal nucleic acid amplification technology
November 2020: Establishment of a Visual Detection Method for Avian Influenza RT-RAA Lateral Flow Test Strips—Hebei Agricultural University
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Avian influenza (AI) is a major disease that poses a significant threat to the healthy development of China’s poultry industry. To establish an efficient and convenient detection method for this disease, we conducted this study. In this assay, primers and probes were designed based on the conserved gene sequence (NP) of the avian influenza virus (AIV). Using reverse transcription recombinase polymerase amplification (RT-RAA), a isothermal amplification technique, we achieved rapid amplification of the NP gene, generating dual-labeled products. The amplified products were then detected using lateral flow dipstick assays (LFDs). By optimizing reaction conditions—including reaction time, temperature, and primer concentration—we developed an RT-RAA-LFD detection method capable of detecting avian influenza virus. The results showed that under 37°C conditions, the target NP gene fragment could be effectively amplified within 24 minutes, and the amplified products could be visually observed with the naked eye using lateral flow dipsticks. The method demonstrated excellent specificity, showing no cross-reactivity with nucleic acids from infectious bronchitis virus (IBV), avian infectious laryngotracheitis virus (AILV), or Newcastle disease virus (NDV). Its sensitivity reached 102 copies/μL, which is 100 times higher than that of conventional PCR. Moreover, the clinical detection agreement rate was 94.8%. This method boasts high specificity, sensitivity, and convenience, making it a feasible approach for field detection of avian influenza virus.
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